MGMT gene promoter methylation assay primer probe system and kit thereof

Abstract

The invention relates to a MGMT gene promoter methylation assay primer probe system and a kit thereof. The system comprises a primer probe group X for determining the DNA quality of a genome, a primer probe group Y for determining methylation conversion ratio and a primer probe group Z for detecting the situation of MGMT gene promoter methylation, wherein the primer probe group X comprises a forward primer a, a reverse primer a and a probe a; the primer probe group Y comprises a forward primer b, a reverse primer b and a probe b; the primer probe group Z comprises a forward primer b, a reverse primer b and a probe c; the 5' terminals of the probe a, the probe b and the probe c are provided with a fluorescence reporter groups; and the 3' terminals of the probe a, the probe b and the probe c are provided with quenching fluorescence groups. The kit is concise in implementation scheme, high in sensitivity and high in accuracy.

Description

technical field [0001] The invention relates to a gene mutation detection product, a detection primer and a detection system used in the product, and belongs to the field of biotechnology. Background technique [0002] o 6 -methylguanine-DNA methyltransferase (O 6 -methylguanine-DNAmethyhransferase, MGMT) is a DNA repair enzyme, which is the only one in human cells that can remove guanine O from DNA 6 Cytotoxic and mutagenic alkylating agent adducts at the site restore damaged guanines, thereby protecting cells from damage by anti-alkylating groups. Promoter methylation is the most common abnormality of MGMT gene, which is closely related to the expression of MGMT protein, leading to the stop of gene transcription and decreased protein expression. [0003] Studies have shown that the higher the methylation rate of the MGMT gene promoter in glioma, the lower the MGMT content, and the higher the malignant degree of the tumor. MGMT gene promoter methylation can also be used...

AI Technical Summary

Problems solved by technology

The MSP method combined with the nested PCR amplification method can improve the specificity of the single MSP method, but the operation time is long, and compared with the MSP method, there are also false positives caused by incomplete bisulfite treatment. The pH value in the modification process is relatively absolute. Accurate, all reagents require fresh configuration, and need to repeatedly explore to find the appropriate reaction time, resulting in cumbersome detection process, and can not achieve quantitative detection, there is a high false positive rate; there is also excessive sulfite treatment, resulting in amplification 3. Methylation Sensitivity Melting Curve Analysis (MS-High Resolution Melting Curve, MS-HRM), which is cumbersome to operate, has a high false positive rate, and is not suitable for the detection of a large number of samples; 4. Fluorescence method (Methylight) , due to the limitations of the probe, there is generally no suitable control system, and the false positive rate is high; 5. Bisulfite sequencing PCR (BSP), which is a PCR combined with Sanger sequencing technology, has accurate results, but the process is cumbersome. Not suitable for high-volume testing, and too expensive to be widely used

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